Bottom Line:
In ARPE-19 cells, RT-PCR and immunofluorescence microscopy showed that Snail mRNA was upregulated by transforming growth factor (TGF)-β and VEGF stimulation.Furthermore, TGF-β induced relocalization of Snail to the nucleus in RPE cells.The current data indicate that Snail is a major transcriptional factor for EMT changes of RPE cells in human CNV.

Mentions:
Next, at 7 days after passage we stimulated ARPE-19 cells with cytokines, TGF-β, TNF-α, VEGF, CTGF, bFGF, and IGF-1, previously found in human CNV samples. Among them, TGF-β and VEGF significantly upregulated Snail mRNA (Figure 5AB). However, the mRNA level of Snail was not changed with stimulation of the other cytokines. Furthermore, fluorescence microscopy depicted the enhanced staining of Snail in the nucleus and cytoplasm of ARPE-19 cells stimulated with TGF-β at 7 days after passage (Figure 5C), indicating a role for TGF-β in the upregulation and nuclear relocalization of Snail in RPE cells. By contrast, VEGF enhanced immunoreactivity of Snail mainly in the cytoplasm, but not in the nucleus (Figure 5C).

Mentions:
Next, at 7 days after passage we stimulated ARPE-19 cells with cytokines, TGF-β, TNF-α, VEGF, CTGF, bFGF, and IGF-1, previously found in human CNV samples. Among them, TGF-β and VEGF significantly upregulated Snail mRNA (Figure 5AB). However, the mRNA level of Snail was not changed with stimulation of the other cytokines. Furthermore, fluorescence microscopy depicted the enhanced staining of Snail in the nucleus and cytoplasm of ARPE-19 cells stimulated with TGF-β at 7 days after passage (Figure 5C), indicating a role for TGF-β in the upregulation and nuclear relocalization of Snail in RPE cells. By contrast, VEGF enhanced immunoreactivity of Snail mainly in the cytoplasm, but not in the nucleus (Figure 5C).

Bottom Line:
In ARPE-19 cells, RT-PCR and immunofluorescence microscopy showed that Snail mRNA was upregulated by transforming growth factor (TGF)-β and VEGF stimulation.Furthermore, TGF-β induced relocalization of Snail to the nucleus in RPE cells.The current data indicate that Snail is a major transcriptional factor for EMT changes of RPE cells in human CNV.